Tyrosinase inhibitory compound

Abstract

To develop a tyrosinase inhibitory compound capable of having skin whitening action, action of enhancing whitening effect, and the like.SOLUTION: A tyrosinase inhibitory compound contains, as an active ingredient, a water-soluble porcine elastin or an elastin-derived peptide being a water-soluble porcine elastin decomposition product (including chemically synthesized compounds equivalent to these). By utilizing the tyrosinase inhibitory compound of this invention in topical skin formulations (e.g., cosmetics), health foods (e.g., dietary supplements), or the like, it can be expected to achieve useful effect of useful skin whitening action and whitening enhancing action caused by compounds derived from biological constituents without toxicity or adverse effects.SELECTED DRAWING: Figure 1

Description

[Technical field]

[0001] The present invention relates to a tyrosinase inhibitor compound, and more particularly to a tyrosinase inhibitor having a skin whitening effect or the like. [Background technology]

[0002] As the proverb goes, "A fair complexion hides seven flaws," fair skin has been an object of envy since ancient times. Even today, fair skin is a subject of interest for people, especially women. The women's information website "mellow" (Non-Patent Document 1) conducted a survey of 200 women over 20 years old about their skin concerns, and nearly 40% of the women answered that fair skin was their number one concern, after controlling pores, acne, wrinkles, dryness, and sagging. Thus, skin whitening has become an essential item for modern women and a major beauty concern.

[0003] Human skin color is made up of three elements: melanin (black), hemoglobin (red), and carotene (yellow), but in reality, skin color is determined mostly by the amount of melanin. The cells that produce melanin are called melanocytes, and in humans, they are located in the basal layer, the lowest layer of the epidermis. Tyrosinase exists in cells called melanocytes, is produced in the ribosomes of the rough endoplasmic reticulum, matures by glycosylation in the Golgi-associated endoplasmic reticulum, and is transferred to and placed in sac-shaped intracellular organelles called melanosomes, where it becomes capable of catalyzing melanin production. The cause of age spots is that the black melanin produced by the action of tyrosinase in melanocytes, the pigment cells of the skin, becomes excessive due to ultraviolet rays, and the turnover cycle for excretion from the skin slows down with age, resulting in the accumulation of the black melanin without being properly excreted. To prevent spots, it is important to suppress the excessive production of melanin or to increase the turnover cycle and promote the excretion of melanin.

[0004] Melanin molecules are large biopolymers, and their raw material is tyrosine, a type of amino acid. Among the main enzymes that synthesize melanin, tyrosinase (EC 1.14.18.1) has attracted attention, and substances that inhibit tyrosinase have a whitening effect. Tyrosinase catalyzes two main reactions in the melanin synthesis process. One is the activity of catalyzing the oxidation of monophenols such as L-tyrosine to o-quinones such as L-dopaquinone (monophenolase activity), and the other is the activity of catalyzing the oxidation of diphenols such as L-DOPA to o-quinones (diphenolase activity).

[0005] Traditionally, various tyrosinase inhibitors have been used for skin whitening, such as kojic acid, hydroquinone, and arbutin, which is a compound in which glucose is bound to the phenolic OH group of hydroquinone. [Prior art documents] [Non-patent literature]

[0006] [Non-Patent Document 1] (Women's information site "mellow" published article on November 15, 2022. URL: https: / / arinna.co.jp / mellow / cosmetics-78 / ) DISCLOSURE OF THEINVENTION [Problem to be solved by the invention]

[0007] Conventionally, tyrosine kinase inhibitors, which have been known as representative examples, have many problems. Specifically, hydroquinone and kojic acid are extremely vulnerable to heat and oxidation, particularly in water, and are unstable, causing decomposition and discoloration over time during preparation. On the other hand, although arbutin has improved stability against heat and oxidation, it is not necessarily satisfactory in terms of inhibitory effect, etc., compared with kojic acid and hydroquinone, when comparing the concentration at which the diphenolase activity of human tyrosinase is inhibited by 50% (IC50). As for side effects, short-term use of hydroquinone can cause redness, inflammation, and rashes, while long-term use can cause white spots. Kojic acid can cause contact dermatitis, and animal experiments have suggested carcinogenicity at high doses, so high doses should be avoided in humans. Arbutin is less likely to cause skin irritation than hydroquinone, but there have been some reports of it causing contact dermatitis.

[0008] Against the background described above, the present invention aims to discover tyrosinase inhibitors that can solve the above-mentioned problems observed with conventional tyrosinase inhibitors, focusing on proteins or their degradation products, which are components of the body, and to provide them as topical skin preparations (cosmetics, etc.) or health foods (supplements, etc.). [Means for solving the problem]

[0009] The inventors conducted research and development focusing on elastin and its degradation products. That is, elastin is a protein that is widely distributed in the skin, blood vessels, ligaments, etc. of the body. When water-soluble porcine elastin is ingested, it is sequentially digested by enzymes in the stomach and small intestine, ultimately producing low molecular weight peptides (elastin degradation products) of amino acids, dipeptides, and tripeptides. Among these low molecular weight peptides are peptides with various physiological activities (active peptides), which are absorbed from the intestinal tract, travel in the blood, and are transported to various tissues in the body. For example, they came up with the idea that when the active peptide is delivered to skin tissue, it may reach melanocytes located in the basal layer at the bottom of the epidermis, inhibit tyrosinase, and suppress melanin synthesis.

[0010] In addition, since the stratum corneum acts as a barrier in skin tissue, high molecular weight substances have difficulty penetrating the skin, but low molecular weight substances with a molecular weight of less than 1,000 are said to pass through the skin relatively easily. We therefore came to the conclusion that by applying water-soluble porcine elastin-derived peptides (elastin hydrolysates) with tyrosinase inhibitory properties and a molecular weight of less than 1,000 to the skin, these water-soluble porcine elastin-derived peptides (elastin hydrolysates) could penetrate the skin and reach melanocytes located in the basal layer at the bottom of the epidermis, inhibiting tyrosinase and suppressing melanin synthesis, thereby whitening the skin.

[0011] The inventors conducted intensive research into these possibilities and discovered that water-soluble porcine elastin or peptides derived from water-soluble porcine elastin (elastin hydrolysates) have an inhibitory effect on tyrosinase, thereby completing the invention.

[0012] The present invention comprises the following configurations. [1] A tyrosinase inhibitor compound characterized by having as an active ingredient water-soluble porcine elastin or a peptide derived from water-soluble porcine elastin (elastin hydrolysate) (including chemically synthesized compounds equivalent thereto).

[0013] [2] The tyrosinase inhibitor compound according to [1], wherein the water-soluble porcine elastin has a molecular weight of less than 300,000, preferably less than 100,000, and particularly preferably has an average molecular weight of 20,000 to less than 30,000. [3] The tyrosinase inhibitor compound according to [1], wherein the molecular weight of the water-soluble porcine elastin-derived peptide (elastin hydrolyzate) is from 100 to less than 2,000, preferably from 100 to less than 1,000. [4] The tyrosinase inhibitor compound described in [3], wherein the water-soluble porcine elastin-derived peptide (elastin degradation product) is represented by any of a dipeptide, tripeptide, tetrapeptide, pentapeptide, hexapeptide, heptapeptide, octapeptide, nonapeptide, decapeptide, undecapeptide, or dodecapeptide contained in the sequence VGVAPGVGVAPGVGVAPGVGVAPG, which is a repeating sequence in porcine tropoelastin (porcine elastin precursor). [5] The tyrosinase inhibitor compound according to [3], wherein the water-soluble porcine elastin-derived peptide (elastin degradation product) is represented by any one of the tripeptides X1X2P, GX1G or AX1A (X1 or X2 = any one of the amino acids represented by A, G, S, V, L, T, I, C, M, D, N, E, Q, H, K, R, F, Y, W, or P) contained in porcine tropoelastin (porcine elastin precursor).

[0014] [6] The tyrosinase inhibitor compound described in [3], wherein the water-soluble porcine elastin-derived peptide (elastin hydrolyzate) is a peptide having a molecular weight of less than 1,000 and including any one or more of SEQ ID NOs: 4 to 21. [7] The tyrosinase inhibitor compound according to [3], wherein the water-soluble porcine elastin-derived peptide (elastin hydrolysate) is represented by sequence numbers 4, 5, 7, 8, 9 and 10.

[0015] [8] A pharmaceutical, topical skin preparation (e.g., cosmetics) or health food (e.g., supplements), preferably a topical skin preparation (e.g., cosmetics) or health food (e.g., supplements), which contains a tyrosinase inhibitor compound of [1] to [7] as an active ingredient and is characterized by a whitening effect or an enhanced whitening effect due to the tyrosinase inhibitor compound inhibiting tyrosinase. Effect of the Invention

[0016] The present invention makes it possible to provide useful tyrosinase inhibitor compounds. By using the tyrosinase inhibitor compound of the present invention as a skin external preparation (e.g., cosmetics) or health food (e.g., supplements), etc., it is expected that a whitening effect will be achieved by using a useful compound derived from a biological component that is easily soluble in water and has no toxicity or side effects, unlike conventional skin external preparations (e.g., cosmetics) or health food (e.g., supplements). [Brief description of the drawings]

[0017] [Figure 1] FIG. 1 shows the results of investigating the tyrosinase inhibition rate (%) and toxicity of water-soluble porcine elastin-derived peptides (elastin hydrolysates). [Diagram 2] This graph shows the results of calculating the IC50 and water solubility of water-soluble porcine elastin and water-soluble porcine elastin-derived peptides (elastin hydrolysates). [Diagram 3] Diagram showing IC50 and water solubility of existing compounds DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0018] The tyrosinase inhibitor compound of the present invention will now be described.

[0019] The tyrosinase inhibitory compound of the present invention is characterized by having as an active ingredient water-soluble porcine elastin or a peptide derived from water-soluble porcine elastin (elastin degradation product) (including a chemically synthesized compound corresponding thereto). That is, the compound exerts a tyrosinase inhibitory effect by using as an active ingredient porcine tropoelastin (porcine elastin precursor) shown in SEQ ID NO: 1 or a peptide derived therefrom (degradation product). The molecular weight of the water-soluble porcine elastin can typically be less than 300,000, preferably less than 100,000, and most preferably an average molecular weight of 20,000 to less than 30,000. Furthermore, the molecular weight of the water-soluble porcine elastin-derived peptide (elastin degradation product) can typically be from 100 to less than 2,000, and preferably from 100 to less than 1,000.

[0020] When the tyrosinase inhibitor compound of the present invention is produced by decomposing elastin, it can be produced by a known method (WO 2006 / 046626). That is, biological tissues such as aorta and ligament obtained from mammals such as pigs, horses, cows, and sheep are used as raw materials, but biological tissues such as avian arterial blood vessels and fish arterial bulbs may also be used as raw materials. First, these biological tissues are degreased, homogenized, etc., and then, collagen and the like are removed from the fragmented sample using an alkaline solution, etc., to obtain insoluble elastin. Furthermore, the insoluble elastin is hydrolyzed with an acid or alkali, etc., and the treated solution is separated to obtain water-soluble elastin. The water-soluble elastin is treated with an enzyme treatment, etc., and the resulting treated solution is subjected to membrane separation or column purification by gel filtration, etc., to obtain elastin-derived peptides (elastin degradation products) consisting of a fraction with a molecular weight of 100 to less than 2,000.

[0021] The tyrosinase inhibitory compound of the present invention can also be chemically synthesized. That is, since the tyrosinase inhibitory compound of the present invention is configured as a peptide, it can be chemically synthesized using a peptide synthesizer or the like.

[0022] In a preferred embodiment of the water-soluble porcine elastin-derived peptide (elastin degradation product), the water-soluble porcine elastin-derived peptide (elastin degradation product) can be a compound having a molecular weight of less than 1,000 and represented by any of a dipeptide, tripeptide, tetrapeptide, pentapeptide, hexapeptide, heptapeptide, octapeptide, nonapeptide, decapeptide, undecapeptide, or dodecapeptide contained in the sequence VGVAPGVGVAPGVGVAPGVGVAPG (sequence number 2). The compound shown in SEQ ID NO:2 is represented by one-letter code for amino acids, with the left side representing the N-terminus and the right side representing the C-terminus (hereinafter the same).

[0023] In addition, as another preferred embodiment of the water-soluble porcine elastin-derived peptide (elastin degradation product), the water-soluble porcine elastin-derived peptide (elastin degradation product) can be a compound having a molecular weight of less than 1,000, represented by a tripeptide of X1X2P or GX1G or AX1A (X1 or X2 = an amino acid represented by any of A, G, S, V, L, T, I, C, M, D, N, E, Q, H, K, R, F, Y, W, or P).

[0024] Furthermore, as another preferred embodiment of the water-soluble porcine elastin-derived peptide (elastin degradation product), the water-soluble porcine elastin-derived peptide (elastin degradation product) can be a peptide having a molecular weight of less than 1,000 including any one or more of SEQ ID NOs: 4 to 21, and particularly preferably a peptide represented by any one or more of SEQ ID NOs: 4, 5, 7, 8, 9, and 10. By using these water-soluble porcine elastin-derived peptides (elastin hydrolysates) as preferred embodiments, the design and chemical synthesis of tyrosinase inhibitory compounds becomes easier, and the tyrosinase inhibitory compounds of the present invention can be obtained with high purity and high reproducibility.

[0025] The tyrosinase inhibitor compound of the present invention has the chemical structure as shown above, but is not limited thereto. That is, in consideration of the purpose of the present invention that the compound having the above-mentioned chemical structure exerts a tyrosinase inhibitory effect, in addition to the case where the compound itself exerts a tyrosinase inhibitory effect, the compound may be a so-called DDS (Drug Delivery System) compound, that is, a PDS (Particle Delivery System) compound, which becomes the compound structure of the present invention through various metabolisms in the body and exerts a tyrosinase inhibitory effect. In this sense, the tyrosinase inhibitor compound of the present invention may be partially chemically modified.

[0026] The tyrosinase inhibitory compound of the present invention can be used in various applications such as medicines, health foods (supplements, etc.), and skin external preparations (cosmetics, etc.) without any particular limitation as long as it can exert a tyrosinase inhibitory effect. Preferred applications include health foods (supplements, etc.), skin external preparations (cosmetics, etc.), and the like. The dosage forms of health foods (supplements, etc.) can be, for example, tablets, powders, granules, capsules, injections, oral liquids, syrups, and the like. The dosage forms of skin external preparations (cosmetics, etc.) can be, for example, powders, balms, creams, liquids, and the like.

[0027] Preferred applications of the tyrosinase inhibitor compound of the present invention include skin whitening applications. In other words, when a human ingests the tyrosinase inhibitor compound of the present invention or applies it to the skin, it inhibits tyrosinase in melanocytes located in the basal layer of skin tissue, thereby suppressing melanin synthesis, thereby contributing to a whitening effect. In such an embodiment, the tyrosinase inhibitor compound of the present invention can be used as an external skin preparation (cosmetics, etc.) or health food (supplements, etc.) that has the characteristic of having a skin whitening effect by using the tyrosinase inhibitor compound as an active ingredient and inhibiting tyrosinase. In a different embodiment, the tyrosinase inhibitor compound of the present invention can be formulated as an external skin preparation (such as cosmetics) or health food (such as supplements) that contains the tyrosinase inhibitor compound of the present invention as an active ingredient and that enhances the skin whitening effect by the tyrosinase inhibitor compound inhibiting tyrosinase.

[0028] The tyrosinase inhibitory compound of the present invention can contain various additives. That is, when the tyrosinase inhibitory compound of the present invention is used as an active ingredient in a composition for external use on the skin (such as cosmetics) or health food (such as supplements), appropriate additives can be selected depending on the dosage form. Such additives include, for example, excipients, stabilizers, antioxidants, pH adjusters, emulsifiers, solubilizers, isotonicity agents and the like.

[0029] Examples of excipients include sugar alcohols such as D-sorbitol, mannitol, xylitol, and propylene glycol, sugars such as glucose, sucrose, lactose, and fructose, crystalline cellulose, calcium hydrogen phosphate, wheat starch, rice starch, corn starch, potato starch, dextrin, β-cyclodextrin, light anhydrous silicic acid, titanium oxide, magnesium aluminometasilicate, talc, kaolin, olive oil, gelatin, casein, and pectin.

[0030] Examples of stabilizers include pectin, dextran, glycerin, polysaccharides (guar gum, gum arabic, xanthan gum, gellan gum, sclerogum, tamarind seed gum), starches (hydroxypropyl starch, phosphate monoesterified phosphate cross-linked starch), agar, gelatin, and methylcellulose.

[0031] Examples of antioxidants include L-ascorbic acids (L-ascorbic acid, L-calcium ascorbate, L-ascorbyl stearate, L-ascorbyl palmitate), erythorbic acid, butylhydroxyanisole, γ-oryzanol, catechin, licorice oil extract, quercetin, citric acid, glyceryl monocitrate, tocotrienol, tocopherols (d-α-tocopherol, d-γ-tocopherol, d-δ-tocopherol), lecithin, ferulic acid, Melaleuca essential oil, sunflower seed extract, grape seed extract, propolis extract, ginkgo extract, bayberry extract, and rosemary extract.

[0032] Examples of pH adjusters that can be used include citric acid, gluconic acid, succinic acid, malic acid, phosphoric acid, maleic acid, tartaric acid, lactic acid, calcium lactate, ammonium acetate, meglumine, and glucono-δ-lactone.

[0033] Examples of the emulsifier include surfactants such as stearyl triethanolamine, sodium lauryl sulfate, lauryl aminopropionic acid, lecithin, benzalkonium chloride, benzethonium chloride, and glycerin monostearate; hydrophilic polymers such as polyvinyl alcohol, polyvinylpyrrolidone, sodium carboxymethylcellulose, methylcellulose, hydroxymethylcellulose, hydroxyethylcellulose, and hydroxypropylcellulose; Examples of such waxes include shellac wax, beeswax, carnauba wax, spermaceti wax, lanolin, liquid lanolin, reduced lanolin, hard lanolin, cyclic lanolin, lanolin wax, candelilla wax, Japan wax, montan wax, shellac wax, and rice wax; sucrose fatty acid esters, sorbitan fatty acid esters, propylene glycol fatty acid esters, glycerin fatty acid esters, calcium stearoyl lactylate, calcium citrate, calcium phosphate salts, soybean saponin, tea seed saponin, beet saponin, sphingolipids, tomato glycolipids, plant sterols, plant lecithin, enzymatically decomposed lecithin, egg yolk lecithin, yucca foam extract, barley husk extract, and quillaja extract.

[0034] Examples of solubilizing agents include macrogol 1500, macrogol 4000, macrogol 6000, macrogol 20000, lauromacrogol, polyoxyl 40 stearate, polysorbate 80, polyoxyethylene hydrogenated castor oil 60, D-mannitol, sodium citrate, anhydrous citric acid, tartaric acid, sodium hydrogen sulfite, and arginine.

[0035] Examples of isotonic agents include glycerin, propylene glycol, sodium chloride, potassium chloride, sodium dihydrogen phosphate, potassium dihydrogen phosphate, citric acid, sodium citrate, sorbitol, mannitol, glucose, and boric acid. EXAMPLES

[0036] The tyrosinase inhibitor compounds of the present invention will be further illustrated by means of experimental examples.

[0037] <<Experimental Example 1, Tyrosinase Inhibition Test by Water-Soluble Porcine Elastin and Water-Soluble Porcine Elastin-Derived Peptides (Elastin Decomposition Products)>> <Experimental Overview> 1.The experiment was performed according to the protocol of the Human Tyrosinase Inhibitor Screening Kit (Diphenolase activity) (Sakulab Science, Japan). The enzyme (tyrosinase) used was of human origin. The substrate was L-DOPA. The inhibitor control was 10 mM Kojic acid. The samples were water-soluble porcine elastin or water-soluble porcine elastin-derived peptides (elastin digests). 2. Enzyme solution was prepared by adding 5μL of the enzyme provided with the kit and 15μL of the assay buffer provided with the kit per well. Next, substrate solution was prepared by adding 5μL of the substrate provided with the kit and 55μL of the assay buffer provided with the kit per well. 20μL of assay buffer (control), inhibitor control, and sample were added to each well of the microplate provided with the kit. Next, 20μL of enzyme solution was added to each well, and after gentle mixing, the mixture was incubated in an incubator (25-30℃) for 10 minutes. Next, 60 L of substrate solution was added to each well, and after gentle mixing, the enzyme reaction was started in an incubator (25-30°C). The absorbance (495 nm) of each well was measured using a plate reader 30 minutes and 60 minutes after the start of the enzyme reaction. The inhibition rate was calculated as follows. △(Control) = absorbance value at 60 minutes (Control) - absorbance value at 30 minutes (Control) △(Sample) = absorbance value at 60 minutes (Sample) - absorbance value at 30 minutes (Sample) If △(Control) > △(Sample): Inhibition rate (%) = △(Control) - △(Sample) / △(Control) X 100 3. The tyrosinase inhibitory activity was examined using a total of 19 compounds, including compounds No. 1 to No. 18 shown in Figure 1 and water-soluble porcine elastin shown in Figure 2.

[0038] 4. Compound numbers 1 to 18 are derived from the following: That is, all of the compounds are derived from porcine tropoelastin (porcine elastin precursor) shown in SEQ ID NO:1. (1) Compounds Nos. 1 to 3, 5, 7, 10, and 14 to 18 are derived from the VGVAPG repeat sequence (VGVAPGVGVAPGVGVAPGVGVAPG, SEQ ID NO: 2) of porcine tropoelastin (porcine elastin precursor) from the 496th Val residue to the 519th Gly residue. (2) Compound No. 13 is a PRP sequence derived from the 692nd Pro residue to the 694th Pro residue of porcine tropoelastin (porcine elastin precursor). (3) Compound No. 11 has a VSP sequence derived from the amino acid residues from the 370th Val residue to the 372nd Pro residue and from the 659th Val residue to the 661st Pro residue of porcine tropoelastin (porcine elastin precursor). (4) Compound No. 12 is a TRP sequence derived from the 682nd Thr residue to the 684th Pro residue of amino acid residues of porcine tropoelastin (porcine elastin precursor). (5) Compound No. 8 has a GAP sequence derived from the amino acid residues of porcine tropoelastin (porcine elastin precursor) from the 249th Gly residue to the 251st Pro residue, from the 283rd Gly residue to the 285th Pro residue, from the 306th Gly residue to the 308th Pro residue, from the 468th Gly residue to the 470th Pro residue, and from the 531st Gly residue to the 533rd Pro residue. (6) Compound No. 6 is a sequence contained in the amino acid residues of porcine tropoelastin (porcine elastin precursor) between the 73rd Ala residue and the 76th Ala residue, between the 235th Ala residue and the 246th Ala residue, between the 286th Ala residue and the 295th Ala residue, between the 373rd Ala residue and the 383rd Ala residue, between the 427th Ala residue and the 434th Ala residue, between the 471st Ala residue and the 482nd Ala residue, between the 539th Ala residue and the 544th Ala residue, between the 589th Ala residue and the 594th Ala residue, between the 623rd Ala residue and the 632nd Ala residue, and between the 662nd Ala residue and the 668th Ala residue. (7) Compound No. 4 has a GPG sequence derived from the 32nd to 34th Gly residues, the 311th to 313th Gly residues, the 487th to 489th Gly residues, and the 523rd to 525th Gly residues of amino acid residues of porcine tropoelastin (porcine elastin precursor). (8) Compound No. 9 has a GLG sequence derived from the amino acid residues of porcine tropoelastin (porcine elastin precursor) from the 21st Gly residue to the 23rd Gly residue, from the 24th Gly residue to the 26th Gly residue, from the 48th Gly residue to the 50th Gly residue, from the 52nd Gly residue to the 54th Gly residue, from the 86th Gly residue to the 88th Gly residue, from the 95th Gly residue to the 97th Gly residue, from the 485th Gly residue to the 487th Gly residue, from the 641st Gly residue to the 643rd Gly residue, and from the 709th Gly residue to the 711th Gly residue.

[0039] 5. The results of tyrosinase inhibition rate and toxicity are shown in Figure 1. (1) Compound 12 (SEQ ID NO: 10), compound 13 (SEQ ID NO: 9), compound 14 (SEQ ID NO: 8), compound 15 (SEQ ID NO: 7), compound 16 (SEQ ID NO: 5) and compound 18 (SEQ ID NO: 4) have excellent tyrosinase inhibitory activity, and all of them showed inhibition rates of 81% or more at a peptide concentration of 40 mM. (2) In particular, compound 14 (SEQ ID NO: 8) showed an inhibition rate of 97% or more, and compound 18 (SEQ ID NO: 4) showed an inhibition rate of 100%. (3) The toxicity of compounds 1 to 18 was examined using the online ToxinPred tool and was found to be non-toxic.

[0040] 6. IC 50 The results of investigating the solubility in water are shown in FIG. (1) Compound 13 (SEQ ID NO: 9), compound 15 (SEQ ID NO: 7), and compound 18 (SEQ ID NO: 4) had relatively low IC values ​​of 18 mM, 15 mM, and 16 mM, respectively. 50 From this, it was considered that Compound 13 (SEQ ID NO: 9), Compound 15 (SEQ ID NO: 7) and Compound 18 (SEQ ID NO: 4) have excellent inhibitory activity against tyrosinase. (2) Furthermore, compound 12 (SEQ ID NO: 10), compound 14 (SEQ ID NO: 8), and compound 16 (SEQ ID NO: 5) were also considered to have inhibitory activity against tyrosinase. (3) In addition, the average molecular weight of water-soluble porcine elastin is usually 20,000 Da to 30,000 Da. Therefore, assuming that the average molecular weight of water-soluble porcine elastin is 25,000 Da, the IC 50 The IC value for Compound 13, Compound 15, and Compound 18 is 0.088 mM. 50 This value was much smaller than that of the control, suggesting that water-soluble porcine elastin has excellent inhibitory activity. (3) The water solubility was calculated using MOE (Molecular Operating Environment), an integrated computational chemistry system that supports drug discovery modality research. It was shown that VAPG (sequence number 8), TRP (sequence number 10), VGVAPG (sequence number 7), and VAPGVG (sequence number 5) all have high water solubility and are easily soluble in water.

[0041] 7. IC of existing tyrosinase inhibitor compounds 50 and its solubility in water is shown in FIG. (1) Arbutin has a lower IC than the water-soluble porcine elastin-derived peptide (elastin hydrolysate) shown in Figure 2. 50 value (high inhibitory activity), but was higher than the IC value of 0.088 mM (88 μM) for water-soluble porcine elastin assuming an average molecular weight of 25,000 Da. 50 values ​​(low inhibitory activity). (2) This is thought to be because when water-soluble porcine elastin is ingested, the numerous water-soluble elastin-derived peptides (compounds 1 to 18), which are produced as elastin degradation products when the elastin is digested and absorbed in the body, act synergistically to exert a greater tyrosinase inhibitory effect than arbutin alone. (3) Regarding the solubility in water, which was determined using MOE (Molecular Operating Environment), an integrated computational chemistry system that supports drug discovery modality research, arbutin has the disadvantage of being poorly soluble in water. 8. These results indicate that water-soluble porcine elastin or peptides derived from water-soluble porcine elastin (elastin degradation products) inhibit tyrosinase. In particular, compounds 15 and 18 had IC 50 The inhibitory rates at a peptide concentration of 40 mM were 90% and 100%, respectively, which were excellent results, and it was found that these compounds are useful as tyrosinase inhibitors. 9. The VGVAPG sequence of compound 15 (SEQ ID NO: 7), which has strong inhibitory activity, occurs as four consecutive repeats in the sequence from positions 496 to 519 (corresponding to SEQ ID NO: 2) in porcine tropoelastin (porcine elastin precursor). Similarly, the VAPGVGVAPGVG sequence of compound 18 (SEQ ID NO: 4), which has strong inhibitory activity, occurs as two consecutive repeats in the sequence from positions 492 to 515 (corresponding to SEQ ID NO: 3) in porcine tropoelastin (porcine elastin precursor). 10. Furthermore, the VAPGVG sequence of compound 16 (SEQ ID NO: 5) and the VAPG sequence of compound 14 (SEQ ID NO: 8), which exhibit inhibitory activity, exist as four consecutive repeats and four discontinuous repeats, respectively, in the above SEQ ID NO: 3. From the presence of such repeats, it was inferred that compound 14 (SEQ ID NO: 8), compound 15 (SEQ ID NO: 7), compound 16 (SEQ ID NO: 5) and compound 18 (SEQ ID NO: 4) are useful for tyrosinase inhibition.

[0042] 11. Based on these experimental results, it is anticipated that ingestion or application to the skin of water-soluble porcine elastin or peptides derived from water-soluble porcine elastin (elastin hydrolysates) may have a skin whitening effect in clinical trials.

Claims

1. A tyrosinase inhibitory compound characterized in that it contains as an active ingredient water-soluble porcine elastin or an elastin-derived peptide (including a chemically synthesized compound equivalent thereto) which is a hydrolysis product of water-soluble porcine elastin.

2. 2. The tyrosinase inhibitor compound according to claim 1, wherein the water-soluble porcine elastin has a molecular weight of less than 300,000, preferably less than 100,000, and particularly preferably has an average molecular weight of from 20,000 to less than 30,000.

3. 2. The tyrosinase inhibitor compound according to claim 1, wherein the molecular weight of the elastin-derived peptide, which is a water-soluble porcine elastin degradation product, is from 100 to less than 2,000, preferably from 100 to less than 1,000.

4. The tyrosinase inhibitor compound described in claim 3, wherein the elastin-derived peptide, which is a water-soluble porcine elastin degradation product, is represented by any of a dipeptide, tripeptide, tetrapeptide, pentapeptide, hexapeptide, heptapeptide, octapeptide, nonapeptide, decapeptide, undecapeptide, or dodecapeptide contained in the sequence VGVAPGVGVAPGVGVAPGVGVAPG, which is a repeating sequence in porcine tropoelastin (porcine elastin precursor).

5. The tyrosinase inhibitor compound according to claim 3, wherein the elastin-derived peptide, which is a hydrolyzate of porcine water-soluble elastin, is represented by any one of the tripeptides X1X2P, GX1G or AX1A (X1 or X2=an amino acid represented by any one of A, G, S, V, L, T, I, C, M, D, N, E, Q, H, K, R, F, Y, W, or P) contained in porcine tropoelastin (porcine elastin precursor).

6. The tyrosinase inhibitor compound according to claim 3, wherein the elastin-derived peptide, which is a water-soluble porcine elastin degradation product, is a peptide having a molecular weight of less than 1,000 and including any one or more of SEQ ID NOs: 4 to 21.

7. The tyrosinase inhibitor compound according to claim 3, wherein the elastin-derived peptide, which is a water-soluble porcine elastin degradation product, is represented by any one or more of SEQ ID NOs: 4, 5, 7, 8, 9, and 10.

8. The tyrosinase inhibitor compound according to claims 1 to 7 can be used as an active ingredient to inhibit tyrosinase, thereby enabling the composition to be formulated as a medicine, topical skin preparation (cosmetics, etc.), or health food (supplements, etc.), preferably a topical skin preparation (cosmetics, etc.) or health food (supplements, etc.), characterized by skin whitening action and enhancing the whitening effect.

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